LOCUS       sequence-178335-        4807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ThsR and mCherry constitutively and sfGFP under the PphsA 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1710 to 1744)
  AUTHORS   Daeffler KN, Galley JD, Sheth RU, Ortiz-Velez LC, Bibb CO, Shroyer 
            NF, Britton RA, Tabor JJ
  TITLE     Engineering bacterial thiosulfate and tetrathionate sensors for 
            detecting gut inflammation.
  JOURNAL   Mol Syst Biol. 2017 Apr 3;13(4):923. doi: 10.15252/msb.20167416.
  PUBMED    28373240
REFERENCE   2  (bases 1 to 4807)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Syst 
            Biol."; date: "2017-04-3"; volume: "13(4):923. doi"; pages: " 
            10.15252/msb.20167416"
COMMENT     Sequence Label: pKD237-3a-2
FEATURES             Location/Qualifiers
     source          1..4807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(131..719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(211..230)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(807..901)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(922..1581)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1496..1515
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        1710..1744
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     RBS             1754..1765
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1773..2483
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1918..1936)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2186..2205
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2584..2602
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2623..2657
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     terminator      3321..3392
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3408..3435
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             3901..3912
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3919..4632
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      4659..4730
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4746..4773
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
       61 catgcaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
      121 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
      181 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
      241 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
      301 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
      361 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
      421 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
      481 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
      541 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
      601 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
      661 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
      721 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
      781 ggattttggt catgactagt gcttggattc tcaccaataa aaaacgcccg gcggcaaccg
      841 agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt
      901 ccaagcgagc tcgatatcaa attacgcccc gccctgccac tcatcgcagt actgttgtaa
      961 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
     1021 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc
     1081 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
     1141 ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
     1201 ataacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
     1261 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
     1321 actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt
     1381 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
     1441 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
     1501 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
     1561 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     1621 aaatacgacc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgggccg
     1681 gccaacgtct cattttcgcc agatatcgat tgacggctag ctcagtccta ggtacagtgc
     1741 tagcgaattc attaaagagg agaaaggtac ccatggtgag caagggcgag gaggataaca
     1801 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
     1861 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     1921 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
     1981 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
     2041 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     2101 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     2161 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     2221 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
     2281 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     2341 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     2401 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     2461 gcatggacga gctgtacaag taagctttct aactaaaaag gcctcccaaa tcggggggcc
     2521 ttttttattg ataacaaaac gctcgtctaa gaaaccatta ttatcatgac attaacctat
     2581 aaaaataggc gtatcacgag gccctttcgt cttcacctcg agtttacggc tagctcagtc
     2641 ctaggtacta tgctagcaga aatataaaga acgatctatt tatccgcgta catgcagcag
     2701 caaatcaacg gcccggtcta cctggtggat gatgatgaag ccattatcga ctccatcgat
     2761 tttttgatgg agggctacgg ttacaaactg aactcgttta actgcggcga tcgctttttg
     2821 gcagaagtcg atctcaccca ggcaggatgt gtaattctgg atgcgcgtat gccaggctta
     2881 actggtcctc aggtgcaaca gctgctgagc gacgcgaaaa gcccgcttgc ggtcatcttc
     2941 ctgaccggcc atggcgatgt tccgatggcg gttgatgcgt tcaaaaatgg cgcgttcgat
     3001 ttctttcaaa aacctgtgcc gggtagcttg ctcagtcagt caattgccaa aggcttgact
     3061 tattcaatcg atcaacatct gaaacgtact aaccaagcgt taatcgacac gctctcggaa
     3121 cgcgaagctc aaatttttca actggtgatt gcaggcaaca ccaacaaaca gatggctaac
     3181 gagctttgcg tggctattcg taccattgag gttcaccgta gcaaactgat gaccaaactg
     3241 ggtgttaaca acctggctga actggttaaa ctggcgccgc tgctggcaca taaatccgaa
     3301 taataatcta gaccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     3361 gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc tcaccttcgg
     3421 gtgggccttt ctgcgtttat agtaaatcac tgcataattc gtgtcgctca aggcgcactc
     3481 ccgttctgga taatgttttt tgcgccgaca tcataacggt tctggcaaat attctgaaat
     3541 gagctggtta gctagtcatt caagcattat tatgctgttt tttgaagtga atgtgcggcc
     3601 atctagccgc acattttgca tctaaaacat gcagtcatca gcaaaataat aaacttttcc
     3661 ccaatatgtg gtttaccaca atttacagga attcactcct gtggtggtgc aaatttgaac
     3721 tgtgaattgc ttcacaaacg ccgctatcgc aatgtcagta tgtggtttac cacaatatct
     3781 aatatcactc tgctcaataa caatgatgaa aaccttagga agaagttaat tgtgttaaac
     3841 agttaactag gggctttatc taacgctctc ctaaggacaa ctgtcattgg gagatttaac
     3901 aaagaggaga aatactagat gcgtaaaggc gaagagctgt tcactggtgt cgtccctatt
     3961 ctggtggaac tggatggtga tgtcaacggt cataagtttt ccgtgcgtgg cgagggtgaa
     4021 ggtgacgcaa ctaatggtaa actgacgctg aagttcatct gtactactgg taaactgccg
     4081 gtaccttggc cgactctggt aacgacgctg acttatggtg ttcagtgctt tgctcgttat
     4141 ccggaccata tgaagcagca tgacttcttc aagtccgcca tgccggaagg ctatgtgcag
     4201 gaacgcacga tttcctttaa ggatgacggc acgtacaaaa cgcgtgcgga agtgaaattt
     4261 gaaggcgata ccctggtaaa ccgcattgag ctgaaaggca ttgactttaa agaagacggc
     4321 aatatcctgg gccataagct ggaatacaat tttaacagcc acaatgttta catcaccgcc
     4381 gataaacaaa aaaatggcat taaagcgaat tttaaaattc gccacaacgt ggaggatggc
     4441 agcgtgcagc tggctgatca ctaccagcaa aacactccaa tcggtgatgg tcctgttctg
     4501 ctgccagaca atcactatct gagcacgcaa agcgttctgt ctaaagatcc gaacgagaaa
     4561 cgcgatcata tggttctgct ggagttcgta accgcagcgg gcatcacgca tggtatggat
     4621 gaactgtaca aatgatgata ataatctaga ccaggcatca aataaaacga aaggctcagt
     4681 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc
     4741 acactggctc accttcgggt gggcctttct gcgtttatag gtacccgtgg atcctctaga
     4801 gtcgacc
//
